Classifications

• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D311/00—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings
  • C07D311/02—Heterocyclic compounds containing six-membered rings having one oxygen atom as the only hetero atom, condensed with other rings ortho- or peri-condensed with carbocyclic rings or ring systems
  • C07D311/04—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring
  • C07D311/58—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring other than with oxygen or sulphur atoms in position 2 or 4
  • C07D311/70—Benzo[b]pyrans, not hydrogenated in the carbocyclic ring other than with oxygen or sulphur atoms in position 2 or 4 with two hydrocarbon radicals attached in position 2 and elements other than carbon and hydrogen in position 6
  • C07D311/72—3,4-Dihydro derivatives having in position 2 at least one methyl radical and in position 6 one oxygen atom, e.g. tocopherols
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P35/00—Antineoplastic agents
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00

Definitions

• One approach to finding new anticancer agents is to determine one or more major targets by which alpha-tocopheryl succinate mediated antineoplastic activities in prostate cancer cells and then develop pharmaceutical agents.
• X is selected from the group consisting of oxygen, nitrogen and sulfur
• Ri is selected from the group consisting of hydrogen, alkyl, carboxylic acid, carboxylate, carboxamide, ester and combinations thereof
• R 2 is selected from the group consisting of alkyl, substituted alkyl, carboxylic acid, carboxylate, carboxamide, sulfonyl, sulfonamide and combinations thereof; and derivatives and metabolites thereof.
• Also provided are prevention and/or treatment of a cell proliferative disease comprising in a subject by administering to the subject a pharmacologically effective dose of a compound of formula I.
• pharmaceutical compositions comprising one or more compounds of formula I.
• Figure 1 shows a first synthetic scheme for preparing the compounds described herein.
• Figure 2 shows a second synthetic scheme for preparing the compounds described herein.
• Figure 3 shows a third synthetic scheme for preparing the compounds described herein.
• Figure 4 shows differential sensitivity of PC-3, LNCaP, and Bcl-xL- overexpressing LNCaP (LNCaP/B3) cells to ⁇ -tocopheryl succinate-induced apoptosis.
• Figure 5 shows ⁇ -Tocopheryl succinate blocks Bcl-xL/Bcl-2 function by inhibiting BH3 domain-mediated heterodimerization.
• Figure 6 shows modeled docking of ⁇ -tocopheryl succinate (upper panel)
• Figure 7 shows structures and potency for inhibiting Bak BH3 peptide binding to
• Figure 8 shows mechanistic validation of the antitumor action of TS-I.
• X is selected from the group consisting of oxygen, nitrogen and sulfur
• R 1 is selected from the group consisting of hydrogen, alkyl, carboxylic acid, carboxylate, carboxamide, ester and combinations thereof
• R 2 is selected from the group consisting of alkyl, substituted alkyl, carboxylic acid, carboxylate, carboxamide, sulfonyl, sulfonamide and combinations thereof; and derivatives and metabolites thereof.
• the compounds of formula I are selected from
• X is selected from the group consisting of oxygen, nitrogen and sulfurjR] is selected from the group consisting of hydrogen, alkyl, carboxylic acid, carboxylate, carboxamide, ester and combinations thereof; R 2 is selected from the group consisting of alkyl, substituted alkyl, carboxylic acid, carboxylate, carboxamide, sulfonyl, sulfonamide and combinations thereof; and derivatives and metabolites thereof.
• X is O
• X-R 1 is either hydroxy or carboxylic acid.
• the pharmaceutical composition includes a therapeutically effective amount of one or more of the compounds of formula I in association with an acceptable carrier.
• the pharmaceutical composition includes a therapeutically effective amount of one or more of the compounds of formula I in association with an acceptable carrier and one or more adjuvants.
• the pharmaceutical composition includes a therapeutically effective amount of one or more of the compounds of formula I in association with an acceptable carrier, one or more adjuvants and one or more diluents.
• one or more of the compounds of formula I may be pharmaceutically acceptable salts thereof.
• one or more of the compounds of formula I may be derivatives of formula I.
• treatment includes partial or total destruction of the undesirable proliferating cells with minimal destructive effects on normal cells.
• desired mechanisms of treatment at the cellular include, but are not limited to one or more of apoptosis, cell cycle arrest, cellular differentiation, or DNA synthesis arrest.
• prevention includes either preventing the onset of a clinically evident unwanted cell proliferation altogether or preventing the onset of a preclinically evident stage of unwanted rapid cell proliferation in individuals at risk. Also intended to be encompassed by this definition is the prevention of metastasis of malignant cells or to arrest or reverse the progression of malignant cells. This includes prophylactic treatment of those at risk of developing precancers and cancers.
• the term "subject" for purposes of treatment includes any human or animal subject who has a disorder characterized by unwanted, rapid cell proliferation. Such disorders include, but are not limited to cancers and precancers.
• the subject is any human or animal subject, and preferably is a human subject who is at risk of acquiring a disorder characterized by unwanted, rapid cell proliferation, such as cancer.
• the subject may be at risk due to exposure to carcinogenic agents, being genetically predisposed to disorders characterized by unwanted, rapid cell proliferation, and so on.
• the compounds of the present invention are also useful for veterinary treatment of mammals, including companion animals and farm animals, such as, but not limited to dogs, cats, horses, cows, sheep, and pigs.
• subject means a human.
• proliferative cells proliferating cells
• proliferating cells proliferating cells
• rapidly proliferating cells proliferative cells
• alkyl is used, either alone or with other terms, such as haloalkyl or alkylaryl, it includes C 1 to Ci 0 linear or branched alkyl radicals, examples include methyl, ethyl, propyl, isopropyl, butyl, terU-butyl, and so forth.
• haloalkyl includes Cj to C 1O linear or branched alkyl radicals substituted with one or more halo radicals. Some examples of haloalkyl radicals include trifluoromethyl, 1,2-dichloroethyl, 3-bromopropyl, and so forth.
• halo includes radicals selected from F, Cl, Br, and I.
• Alkyl radical substituents of the present invention may also be substituted with other groups such as azido, for example, azidomethyl, 2-azidoethyl, 3-azidopropyl and so on.
• aryl used alone or in combination with * other terms such as alkylaryl, haloaryl, or haloalkylaryl, includes such aromatic radicals as phenyl, biphenyl, and benzyl, as well as fused aryl radicals such as naphthyl, anthryl, phenanthrenyl, fluorenyl, and indenyl and so forth.
• aryl also encompasses "heteroaryls,” which are aryls that have carbon and one or more heteroatoms. such as O, N, or S in the aromatic ring. Examples of heteroaryls include indolyl, pyrrolyl, and so on.
• “Alkylaryl” or “arylalkyl” refers to alkyl-substituted aryl groups such as butylphenyl, propylphenyl, ethylphenyl, methylphenyl, 3,5-dimethylphenyl, tert- butylphenyl and so forth.
• Haloaryl refers to aryl radicals in which one or more substitutable positions has been substituted with a halo radical, examples include fluorophenyl, 4-chlorophenyl, 2,5-chlorophenyl and so forth.
• Haloalkylaryl refers to aryl radicals that have a haloalkyl substituent. Examples of haloalkylaryls include such radicals as bromomethylphenyl, 4-bromobutylphenyl and so on.
• Carboxyamide refers to the group -CONH 2
• sulfonamide refers to the group -SO 2 NH2.
• compositions of formula I are also included in the family of compounds of formula I.
• pharmaceutically acceptable salts connotes salts commonly used to form alkali metal salts and to form addition salts of free acids or free bases.
• the nature of the salt is not critical, provided that it is pharmaceutically acceptable.
• Suitable pharmaceutically acceptable acid addition salts of compounds of formula I may be prepared from an inorganic acid or from an organic acid. Examples of such inorganic acids are hydrochloric, hydrobromic, hydroiodic, nitric, carbonic, sulfuric, and phosphoric acid.
• Appropriate organic acids may be selected from aliphatic, cyclo aliphatic, aromatic, araliphatic, heterocyclic, carboxylic, and sulfonic classes of organic acids, examples of which include formic, acetic, propionic, succinic, glycolic, gluconic, lactic, malic, tartaric, citric, ascorbic, glucoronic, maleic, fumaric, pyruvic, aspartic, glutamic, benzoic, anthranilic, mesylic, salicylic, /7-hydroxybenzoic, phenylacetic, mandelic, ambonic, pamoic, methanesulfonic, ethanesulfonic, benzenesulfonic, pantothenic, 2-hydroxyethanesulfonic, toluenesulfonic; sulfanilic, cyclohexylaminosulfonic, stearic, algenic, yS-hydroxybutyric, galact
• Suitable pharmaceutically acceptable base addition salts of compounds of formula I include metallic salts made from aluminum, calcium, lithium, magnesium, potassium, sodium, and zinc.
• organic salts made from N,N'-dibenzylethylenediamine, chloroprocaine, choline, diethanolamine, ethylenediamine, meglumine (N-methylglucamine) and procaine may be used form base addition salts of the compounds of formula I. All of these salts may be prepared by conventional means from the corresponding compounds of formula I by reacting, for example, the appropriate acid or base with the compound of formula I.
• compositions for the prevention and/or treatment of undesirable, rapidly proliferating cells, such as for treating, preventing, or delaying the onset of a cancer in a subject in need of such treatment.
• the pharmaceutical composition comprises a therapeutically effective amount of a compound of formula I, or a derivative or pharmaceutically acceptable salt thereof, in association with at least one pharmaceutically acceptable carrier, adjuvant, or diluent (collectively referred to herein as "carrier materials") and, if desired, other active ingredients.
• carrier materials collectively acceptable carrier, adjuvant, or diluent
• active compounds of the present invention may be administered by any suitable route known to those skilled in the art, preferably in the form of a pharmaceutical composition adapted to such a route, and in a dose effective for the treatment intended.
• the administration of the present invention may be for either prevention or treatment purposes.
• the methods and compositions used herein may be used alone or in conjunction with additional therapies known to those skilled in the art in the prevention or treatment of disorders characterized by unwanted, rapid proliferation of cells.
• the methods and compositions described herein may be used as adjunct therapy.
• the apoptosis-inducing compounds of the present invention may be administered alone or in conjunction with other antineoplastic agents or other growth inhibiting agents or other drugs or nutrients.
• antineoplastic agents available in commercial use, in clinical evaluation and in pre-clinical development, which could be selected for treatment of cancers or other disorders characterized by rapid proliferation of cells by combination drug chemotherapy.
• Such antineoplastic agents fall into several major categories, namely, antibiotic- type agents, alkylating agents, antimetabolite agents, hormonal agents, immunological agents, interferon-type agents and a category of miscellaneous agents.
• other antineoplastic agents such as metallomatrix proteases inhibitors (MMP), may be used.
• MMP metallomatrix proteases inhibitors
• Suitable agents which may be used in combination therapy will be recognized by those of skill in the art.
• radioprotective agents known to those of skill in the art may also be used.
• adjunct therapy in defining use of a compound of the present invention and one or more other pharmaceutical agent, is intended to embrace administration of each agent in a sequential manner in a regimen that will provide beneficial effects of the drug combination, and is intended as well to embrace co-administration of these agents in a substantially simultaneous manner, such as in a single formulation having a fixed ratio of these active agents, or in multiple, separate formulations for each agent.
• the pharmaceutical composition may be in the form of, for example, a tablet, capsule, suspension or liquid.
• the pharmaceutical composition is preferably made in the form of a dosage unit containing a particular amount of the active ingredient.
• dosage units are capsules, tablets, powders, granules or a suspension, with conventional additives such as lactose, mannitol, corn starch or potato starch; with binders such as crystalline cellulose, cellulose derivatives, acacia, corn starch or gelatins; with disintegrators such as corn starch, potato starch or sodium carboxymethyl-cellulose; and with lubricants such as talc or magnesium stearate.
• the active ingredient may also be administered by injection as a composition wherein, for example, saline, dextrose or water may be used as a suitable carrier.
• the compound may be combined with a sterile aqueous solution which is preferably isotonic with the blood of the recipient.
• a sterile aqueous solution which is preferably isotonic with the blood of the recipient.
• Such formulations may be prepared by dissolving solid active ingredient in water containing physiologically compatible substances such as sodium chloride, glycine, and the like, and having a buffered pH compatible with physiological conditions to produce an aqueous solution, and rendering said solution sterile.
• the formulations may be present in unit or multi-dose containers such as sealed ampoules or vials.
• the compound may be formulated with acid-stable, base-labile coatings known in the art which begin to dissolve in the high pH small intestine. Formulation to enhance local pharmacologic effects and reduce systemic uptake are preferred.
• Formulations suitable for parenteral administration conveniently comprise a sterile aqueous preparation of the active compound which is preferably made isotonic. Preparations for injections may also be formulated by suspending or emulsifying the compounds in non-aqueous solvent, such as vegetable oil, synthetic aliphatic acid glycerides, esters of higher aliphatic acids or propylene glycol.
• non-aqueous solvent such as vegetable oil, synthetic aliphatic acid glycerides, esters of higher aliphatic acids or propylene glycol.
• the active ingredient may be formulated into suppositories using bases which are solid at room temperature and melt or dissolve at body temperature.
• bases include cocoa butter, glycerinated gelatin, hydrogenated vegetable oil, polyethylene glycols of various molecular weights, and fatty esters of polyethylene stearate.
• the pharmaceutical compositions may contain active ingredient in the range of about 0.1 to 2000 mg, preferably in the range of about 0.5 to 500 mg and most preferably between about 1 and 200 mg.
• the daily dose can be administered in one to four doses per day.
• Cell culture - LNCaP androgen-dependent (p53+/+) and PC-3 androgen- nonresponsive (P53-/-) prostate cancer cells were obtained from the American Type Culture Collection (Manassas, VA). The preparation of the stable Bcl-xL-overexpressing LNCaP clone B3 (LNCaP/B3) was previously described (18).
• PC-3, LNCaP, and LNCaP/B3 cells were maintained in RPMI 1640 supplemented with 10% fetal bovine serum (FBS) at 37 0 C in a humidified incubator containing 5% carbon dioxide.
• Normal human prostate epithelial (PrEC) cells were purchased from Cambrex Bio Science Walkersville, Inc. (East Tutherford, NJ).
• Reagents - ⁇ -Tocopherol, ⁇ -tocopheryl succinate, 2,2,5,7,8-pentamethyl-6- chromanol and other chemical reagents required for the synthesis of various analogues were purchased from Aldrich Sigma (St. Louis, MO) unless otherwise indicated.
• TS-I succinic acid mono-[2-(4,8-dimethyl-nonyl)-2,5,7,8-tetramethyl-chroman-6-yl] ester
• TS-2 succinic acid mono-[2,5,7,8-tetramethyl-2-(4-methyl-pentyl)-chroman-6-yl] ester
• TS-3 succinic acid mono-(2,2,5,7,8-pentamethyl-chroman-6-yl) ester
• TS-4 (2-(4,8-dimethyl-nonyl)- 2,5,7,8-tetramethyl-chroman-6-ol
• TS-5 (3-[2,5,7,8-tetramethyl-2-(4,8-dimethyl-nonyl)- chroman-6-yloxy]propionic acid) will be published elsewhere.
• Rabbit antibodies against Bad, cytochrome c, and mouse anti-Bcl-2 were from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA).
• Mouse monoclonal anti-actin was from ICN Biomedicals, Inc. (Costa Mesa, CA).
• Cell viability analysis The effect of individual test agents on cell viability was assessed by using the MTT assay in 6 to 12 replicates.
• PC3, LNCaP, and B3-LNCaP cells were seeded and incubated in poly-p-lysine-coated, 96- well, flat-bottomed plates in RPMI 1640 medium supplemented with 10% FBS medium for 24 hours.
• PrEC cells were seeded at the recommend density in 96-well, flat-bottomed plates in Prostate Epithelial Cell Medium with growth supplements for 3 days.
• the pellet fraction was recovered, placed on ice, and triturated with 100 ⁇ L of a chilled hypotonic lysis solution [50 mM PIPES- KOH (pH 7.4) containing 220 mM mannitol, 68 mM sucrose, 50 mM KCl, 5 mM EDTA, 2 mM MgCb, 1 mM dithiothreitol, and a mixture of protease inhibitors including 100 ⁇ M 4-(2- aminoethyl)benzenesulfonyl fluoride, 80 nM aprotinin, 5 ⁇ M bestatin, 1.5 ⁇ M E-64 protease inhibitor, 2 ⁇ M leupeptin, and 1 ⁇ M pepstatin A].
• a chilled hypotonic lysis solution [50 mM PIPES- KOH (pH 7.4) containing 220 mM mannitol, 68 mM sucrose, 50 mM KCl, 5 m
• the adherent cells in T-25 or T-75 flasks were scraped, combined with the medium, and centrifuged at 2200 rpm for 10 min.
• the supernatants were recovered, placed on ice, and triturated with 20 to 50 ⁇ L of a chilled lysis buffer (M-PER Mammalian Protein Extraction Reagent; Pierce, Rockford, IL), to which was added 1 % protease inhibitor cocktail (set IQ; EMD Biosciences, Inc., San Diego, CA). After a 30-min incubation on ice, the mixture was centrifuged at 16,100 x g for 3 min.
• M-PER Mammalian Protein Extraction Reagent Pierce, Rockford, IL
• set IQ protease inhibitor cocktail
• the immunocomplex was collected by brief centrifugation, washed four times with 800 ⁇ L of ice-cold radioimmunoprecipitation assay buffer, and suspended in 50 ⁇ L of 2 x SDS sample loading buffer. The suspension was boiled for 10 min, cooled, and briefly centrifuged to remove the beads. Western blot analysis with antibodies against Bak as described above.
• LGA Lamarckian genetic algorithm
• figure 1 shows the synthetic scheme for preparing the compounds described herein.
• Figure 4 shows differential sensitivity of PC-3, LNCaP, and Bcl-xL-overexpressing LNCaP (LNCaP/B3) cells to ⁇ -tocopheryl succinate-induced apoptosis.
• B Evidence of apoptotic death in ⁇ -tocopheryl succinate-treated PC-3 cells.
• FIG. 7 shows structures and potency for inhibiting Bak BH3 peptide binding to Bcl-xL and for suppressing the viability of PC-3 and LNCaP cells for ⁇ -tocopheryl succinate and TS-I - TS-5.
• the general structure of ⁇ -tocopheryl succinate and TS-I - TS-3 and structures of TS-4 and TS-5 are shown at the top.
• N represents the number of the isopranyl units in the aliphatic side chain.
• a-Tocopheryl succinate is an inhibitor of Bcl-xL function
• ⁇ -tocopheryl succinate-mediated apoptosis might involve the modulation of the function of Bcl-xL and/or other Bcl-2 members. Accordingly, we examined this putative link at both transcriptional and posttranscriptional levels. First, we assessed the dose-dependent effect of ⁇ -tocopheryl succinate on the expression of different Bcl-2 family members in PC-3 cells, including Bcl-xL, Bcl-2, Bax, Bak, Bad, and Bid by Western blotting. Fig.
• Bcl-xL ⁇ -Tocopheryl succinate was docked into the Bak peptide-binding site that is located in a hydrophobic cleft bound by the BHl, BH2, and BH3 regions of Bcl-xL. Docking analysis indicates that ⁇ -tocopheryl succinate adopted a unique hairpin-shaped conformation in interacting with this hydrophobic pocket (Fig. 6A). As shown, the carboxylic terminus of the hemisuccinate formed electrostatic interactions and hydrogen bonding with the guanidino side chain of ArglOO.

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• 2007
  • 2007-02-21 AU AU2007217810A patent/AU2007217810B2/en not_active Ceased
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